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Method for enzymatic preparation of glycerophosphorylcholine

A technology for glycerophosphocholine and enzymatic preparation, applied in directions such as fermentation, can solve the problems of high cost, difficult to achieve large-scale production, complex process and the like

Inactive Publication Date: 2015-08-05
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are studies using tetrabutylammonium hydroxide as a catalyst to hydrolyze lecithin to prepare L-α-GPC. The process is complicated, the cost is high, and it is difficult to realize large-scale production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Get the phospholipid mixture of 20g (the content of PC is 85%, the content of LPC is 0.7%, the content of GPC is 0.1%, is obtained by extraction of egg yolk phospholipids), and the buffer solution (0.25M citric acid-NaOH ), add 0.2g of phospholipase Lecitase Ultra (Novozymes product, with phospholipase A1 activity), and add 0.4g of partial glyceride lipase lipase G (Amano company product, diglyceride lipase) simultaneously, and The mixture was homogenized for 1 min, and the reaction mixture was stirred normally at 40 °C for the enzymatic reaction. Take a sample after reacting for 30 minutes, and use liquid chromatography to analyze the enzyme reaction product to obtain a PC content of 38.5%, an LPC content of 4.6%, and a GPC content of 42.8%; react for 1 hour and take a sample, and use liquid chromatography to analyze the enzyme reaction The content of PC obtained from the product was 1.7%, the content of LPC was 1.5%, the content of GPC was 82.6%, and the yield of GPC ...

Embodiment 2

[0030] Get the phospholipid mixture of 20g (the content of PC is 85%, the content of LPC is 0.7%, the content of GPC is 0.1%, is obtained by extraction of egg yolk phospholipids), and the buffer solution (0.25M citric acid-NaOH ), add the phospholipase Lecitase Ultra of 0.2g (Novozymes company product, have phospholipase A1 activity), add the partial glyceride lipase Lipase SMG1 of 0.4g simultaneously, and the mixture is homogeneous 1 minute, the reaction mixture is at 30 The enzymatic reaction was carried out under normal stirring at ℃. After reacting for 30 minutes, take a sample, and use liquid chromatography to analyze the enzyme reaction product to obtain a PC content of 31.2%, an LPC content of 3.4%, and a GPC content of 51.2%; react for 1 hour and take a sample, and use liquid chromatography to analyze the enzyme reaction The content of PC obtained from the product was 1.2%, the content of LPC was 1.1%, the content of GPC was 83.4%, and the yield of GPC was 97.8%.

Embodiment 3

[0032]Get the phospholipid mixture of 20g (the content of PC is 85%, the content of LPC is 0.7%, the content of GPC is 0.1%, is obtained by extraction of egg yolk phospholipids), and the buffer solution (0.25M citric acid-NaOH ), add the phospholipase Lecitase Ultra of 0.4g (Novozymes company product, have phospholipase A1 activity), add the partial glyceride lipase Lipase SMG1 of 0.2g simultaneously, and the homogeneous mixture 1 minute, the reaction mixture is at 30 The enzymatic reaction was carried out under normal stirring at ℃. After reacting for 30 minutes, take a sample, and use liquid chromatography to analyze the enzyme reaction product to obtain a PC content of 18.1%, an LPC content of 23.6%, and a GPC content of 44.1%; react for 1 hour and take a sample, and use liquid chromatography to analyze the enzyme reaction The content of PC obtained from the product was 0.7%, the content of LPC was 3.4%, the content of GPC was 81.7%, and the yield of GPC was 96.1%.

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Abstract

The invention discloses a method for enzymatic preparation of glycerophosphorylcholine. The method comprises the following steps: (1) mixing phosphatidylcholine-containing phospholipid with a hydroxyl donor, phospholipase and partial glyceride lipase, and then conducting hydrolysis or alcoholysis reaction, wherein phospholipase is one or the mixture of two of phospholipase A and phospholipase B; and (2) separating resultants and reclaiming glycerophosphorylcholine. The invention relates to the method in which phospholipase and partial glyceride lipase are together used for enzyme reaction to catalyze phosphatidylcholine in the substrate and remove acyl from fatty acid to prepare glycerophosphorylcholine. When partial glyceride lipase and phospholipase are used together, the conversion rate of phosphatidylcholine, the enzyme reaction rate and the yield of glycerophosphorylcholine are far higher than independent use of phospholipase, the conversion rate of phosphatidylcholine in the substrate can be more than 95%, and the reaction duration can be less than one hour.

Description

technical field [0001] The invention relates to a method for enzymatically preparing glycerol phosphorylcholine. Background technique [0002] L-α-Glycerophosphocholine (CAS No. 28319-77-9) is a naturally occurring substance in nature, and is a water-soluble substance produced by removing two fatty acids from phosphatidylcholine (PC), the main phospholipid component of biological cell membranes . Glycerophosphorylcholine exists widely in organisms and is one of the components of human breast milk and body fluids. In the revision of food and drug classification in 2009, glycerophosphocholine was defined as food. In recent years, scientific research has shown that L-α-glycerophosphorylcholine also has important medical value. It can improve and enhance the cognitive ability of the brain, and has certain effects on the growth and development of adolescents and the improvement of memory. Cognitive damage caused by early Alzheimer's disease can be repaired to some extent. At ...

Claims

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Application Information

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IPC IPC(8): C12P13/00
Inventor 王卫飞王永华杨博严慧玲蓝东明李志刚
Owner SOUTH CHINA UNIV OF TECH
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